RealStart
DNA Polymerase Premix
Ver. L0619
Cat. No.
FYT101-100P
FYT102-100P
RealStart
DNA Polymerase Premix
Volume: 1.25 ml
Table 1. Reaction components for RealStart PCR mixture
Storage: -20°C
Description
RealStart DNA Polymerase premix is an ultra-sensitive and convenient PCR
premix product. It contains 2× concentrated solution of HotStart DNA
polymerase, dNTPs, optimized buffers, and loading dye (optional) needed
for PCR. The only step it takes to perform PCR with RealStart DNA
Polymerase premix is to add DNA template and primers into the reaction
mix. Since special HotStart DNA polymerase in the premix is activated after
heating, it greatly reduces non-specific amplification when working with the
premix at room temperature.
Component
FYT101-100P
FYT102-100P
2X RealStart
DNA Polymerase Premix
1.25 ml
1.25 ml
(w/o loading dye)
(w/ loading dye)
RealStart DNA Polymerase Premix Contents:
Hotstart Taq DNA polymerase
dNTPs mix (including dATP, dCTP, dGTP, dTTP)
7.5 mM MgCl2
Loading dye (including bromphenol blue)
Quality Control
Nuclease activity is not detected after incubation 1 µg lambda/HindIII
DNA with 5 units RealStart DNA polymerase in 50 µl of the supplied
reaction buffer for 18 hours at 37°C.
Procedure
A. Preparation of the PCR Master Mix
1. Prepare a master mix according to Table 1.
2. Mix the master mix thoroughly by pipetting up and down.
Dispense the master mix into PCR tubes or plates.
Volume
12.5 µl
1X
Forward Primer (10 µM)
0.75 µl
0.3 µM
0.75 µl
0.3 µM
Reverse Primer (10 µM)
Template DNA
Final conc.
0.5 -10 µl
ddH2O
variable
Total volume
25 µl
B. Performing PCR
1. Program your instrument according to Figure 1.
2. Place the PCR tubes or PCR plates in the thermo cycler and start
the cycling program.
94 °C
94°C
15 min
30 sec
72°C
72°C
1 min/kb
7 min
Primer Tm -5 °C
30 sec
4 °C
∞
35-45 cycles
Unit Definition
One unit is defined as the amount of enzyme that will incorporate 10
nmol of dNTPs into acid-insoluble material in 30 minutes at 72 °C.
Component
RealStart DNA Polymerase Premix
Figure 1. RealStart PCR cycling conditions
Features and Benefits
Less Chance of Contamination – Reduce non-specific amplification
caused by mispriming events that occur during setup and initial
temperature increase.
High Sensitivity – The sensitivity level is equivalent to that of real-time
PCR.
Convenient –An excellent tool when working with large quantities of
samples.
Work well with T&A™ Cloning Kit (FYC001-20P)
Note
For research use only. Not for use in diagnostic or therapeutic procedures.